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Peroxyoxalate Chemiluminescence Assay of Hydrogen Peroxide and Glucose Using 2,4,6,8-Tetrathiomorpholuiopyrimido[5,4-d]-pyrimidine as a Fluorescent Component

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Abstract

Peroxyoxalate chemiluminescence (CL) assay of hydrogen peroxide (H2O2) or glucose was developed by using 2,4,6,8-tetrathiomorpholinopyrimido[5,4-d]pyrimidine as a fluorescent component and bis(2,4,6-trichlorophenyl)oxalate (TCPO) as an oxalate. Linear relationships between CL intensity and final concentration of H2O2 from 10-8 to 10-4 M were obtained. The detection limit at the ratio of CL intensities for sample and blank (S/B) of 3 was 10 nM. The precision for five replicate measurements at 10-5 and 10-6 M of H2O2 were 17.6 and 15.7% of relative standard deviations, respectively. α-D-Glucose was transformed to β-D-glucose with mutarotase and converted to H2O2 and D-gluconic acid with glucose oxidase, which was detected by using peroxyoxalate CL reaction. A linear calibration graph was obtained up to 1.5×10-4 M of glucose solution. The method was applied to the assay of glucose in human serum. The recovery was 98.2% (N=4). The method correlated well with the conventional colorimetric method (r=0.968).

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Authors and Affiliations

  1. School of Pharmaceutical Sciences, Nagasaki University, Bunkyo-machi, Nagasaki, 852, Japan

    Kenichiro Nakashima, Kouichi Maki, Shinki Kawaguchi & Shuzo Akiyama

  2. Branch Hospital Pharmacy, University of Tokyo, Mejirodai, Bunkyo, Tokyo, 112, Japan

    Yukie Tsukamoto & Kazuhiro Imai

Authors
  1. Kenichiro Nakashima
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  2. Kouichi Maki
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  3. Shinki Kawaguchi
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  4. Shuzo Akiyama
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  5. Yukie Tsukamoto
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  6. Kazuhiro Imai
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Nakashima, K., Maki, K., Kawaguchi, S. et al. Peroxyoxalate Chemiluminescence Assay of Hydrogen Peroxide and Glucose Using 2,4,6,8-Tetrathiomorpholuiopyrimido[5,4-d]-pyrimidine as a Fluorescent Component. ANAL. SCI. 7, 709–713 (1991). https://doi.org/10.2116/analsci.7.709

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  • DOI: https://doi.org/10.2116/analsci.7.709

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